U.S. patent application number 11/184689 was filed with the patent office on 2006-02-02 for benzoic acid derivatives as non nucleoside reverse transcriptase inhibitors.
This patent application is currently assigned to Boehringer Ingelheim International GmbH. Invention is credited to Jeffrey O'Meara, Bruno Simoneau, Christiane Yoakim.
Application Number | 20060025480 11/184689 |
Document ID | / |
Family ID | 35786846 |
Filed Date | 2006-02-02 |
United States Patent
Application |
20060025480 |
Kind Code |
A1 |
Simoneau; Bruno ; et
al. |
February 2, 2006 |
Benzoic acid derivatives as non nucleoside reverse transcriptase
inhibitors
Abstract
Compounds of formula (I): ##STR1## wherein R.sup.1, R.sup.2,
R.sup.3, R.sup.4, R.sup.5 and R.sup.6 are as defined herein. The
compounds are useful as reverse transcriptase inhibitors against
HIV. In particular, the compounds are active against wild type and
single or double mutant strains of HIV.
Inventors: |
Simoneau; Bruno; (Laval,
CA) ; O'Meara; Jeffrey; (Boisbriand, CA) ;
Yoakim; Christiane; (Laval, CA) |
Correspondence
Address: |
MICHAEL P. MORRIS;BOEHRINGER INGELHEIM CORPORATION
900 RIDGEBURY RD
P O BOX 368
RIDGEFIELD
CT
06877-0368
US
|
Assignee: |
Boehringer Ingelheim International
GmbH
Ingelheim
DE
|
Family ID: |
35786846 |
Appl. No.: |
11/184689 |
Filed: |
July 19, 2005 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
60598227 |
Aug 2, 2004 |
|
|
|
Current U.S.
Class: |
514/563 ;
562/450 |
Current CPC
Class: |
C07C 235/24 20130101;
C07C 235/68 20130101; C07C 255/56 20130101; A61P 31/18
20180101 |
Class at
Publication: |
514/563 ;
562/450 |
International
Class: |
A61K 31/195 20060101
A61K031/195; C07C 235/80 20060101 C07C235/80 |
Claims
1. A compound of the formula (I) ##STR23## wherein, R.sup.1 and
R.sup.2 are each independently selected from H, halo, cyano,
(C.sub.1-4)alkyl, (C.sub.1-4)haloalkyl, (C.sub.2-4)alkenyl,
(C.sub.2-4)alkynyl and (C.sub.3-6)cycloalkyl; with the proviso that
when R.sup.1 is H, R.sup.2 cannot be H; R.sup.3 is halo; R.sup.4 is
selected from (C.sub.1-4)alkyl, halo and nitro; and R.sup.5 and
R.sup.6 are each independently selected from H, halo and
(C.sub.1-4)alkyl; or a salt thereof.
2. A compound according to claim 1 wherein R.sup.1 and R.sup.2 are
each independently selected from H, halo, cyano, (C.sub.1-4)alkyl,
(C.sub.1-4)haloalkyl, and (C.sub.3-6)cycloalkyl; with the proviso
that when R.sup.1 is H, R.sup.2 cannot be H.
3. A compound according to claim 2 wherein R.sup.1 and R.sup.2 are
each independently selected from H, fluoro, chloro, bromo, cyano,
trifluoromethyl and cyclopropyl; with the proviso that when R.sup.1
is H, R.sup.2 cannot be H.
4. A compound according to claim 1 wherein R.sup.3 is chloro.
5. A compound according to claim 1 wherein R.sup.4 is selected from
chloro, bromo, nitro and methyl.
6. A compound according to claim 5 wherein R.sup.4 is chloro, bromo
or methyl.
7. A compound according to claim 1 wherein R.sup.5 is H, fluoro,
chloro or methyl.
8. A compound according to claim 1 wherein R.sup.6 is H or
fluoro.
9. A compound according to claim 1 wherein R.sup.1 and R.sup.2 are
each independently selected from H, halo, cyano, (C.sub.1-4)alkyl,
(C.sub.2-4)alkenyl, (C.sub.2-4)alkynyl and (C.sub.3-6)cycloalkyl;
wherein said (C.sub.1-4)alkyl is optionally substituted with one to
three halo substituents; with the proviso that when R.sup.1 is H,
R.sup.2 cannot be H; R.sup.3 is halo; R.sup.4 is selected from
(C.sub.1-4)alkyl, halo and nitro; R.sup.5 is selected from H and
halo; and R.sup.6 is H; or a pharmaceutically acceptable salt
thereof.
10. A compound according to claim 1 wherein R.sup.1 and R.sup.2 are
each independently selected from H, halo, cyano, (C.sub.1-4)alkyl,
(C.sub.1-4)haloalkyl, and (C.sub.3-6)cycloalkyl; with the proviso
that when R.sup.1 is H, R.sup.2 cannot be H; R.sup.3 is chloro;
R.sup.4 is chloro, bromo, nitro or methyl; R.sup.5 is H, fluoro,
chloro or methyl; and R.sup.6 is H or fluoro.
11. A compound according to claim 1 wherein R.sup.1 and R.sup.2 are
each independently selected from H, fluoro, chloro, bromo, cyano,
trifluoromethyl and cyclopropyl; with the proviso that when R.sup.1
is H, R.sup.2 cannot be H; R.sup.3 is chloro; R.sup.4 is chloro,
bromo or methyl; R.sup.5 is H, fluoro, chloro or methyl; and
R.sup.6 is H or fluoro.
12. A compound of the formula ##STR24## wherein R.sup.1, R.sup.2,
R.sup.4, R.sup.5 and R.sup.6 are as defined in the table
immediately below. TABLE-US-00003 Cpd R.sup.1 R.sup.2 R.sup.4
R.sup.5 R.sup.6 1001 F CF.sub.3 Me H H; 1002 F CF.sub.3 Me F H;
1003 F CN Me F H; 1004 F ##STR25## Me H H; 1005 F Cl Me H H; 1006 H
CN Me H H; 1007 F CN Me H H; 1008 F CF.sub.3 Cl H H; 1009 F CN Cl H
H; 1010 F CF.sub.3 Cl F H; 1011 F ##STR26## Me F H; 1012 F Br Me F
H; 1013 F Cl Me F H; 1014 F Br Me H H; 1015 H Br Me H H; 1016 H Br
Me F H; 1017 H CN Me F H; 1018 Cl CN Me H H; 1019 F ##STR27## Br H
H; 1020 F CF.sub.3 Br H H; 1021 Cl CN Me F H; 1022 F CF.sub.3 Cl Cl
H; 1023 F CF.sub.3 Me Me H; 1024 F CF.sub.3 Me Cl H; or 1025 F
CF.sub.3 Me H F.
13. A pharmaceutical composition, comprising a compound according
to claim 1, or a pharmaceutically acceptable salt thereof, and one
or more pharmaceutically acceptable carriers.
14. A composition according to claim 13 additionally comprising one
or more other antiretroviral drugs.
15. A composition according to claim 14 wherein the one or more
other antiretroviral drugs are selected from the group consisting
of NRTIs, NNRTIs, protease inhibitors, entry inhibitors, integrase
inhibitors, TAT inhibitors, maturation inhibitors, immunomodulating
agents, antifungal agents and antibacterial agents.
16. A method of treating HIV infection in a human which comprises
administering a therapeutically effective amount of a compound
according to claim 1 or a pharmaceutically acceptable salt thereof.
Description
RELATED APPLICATIONS
[0001] This application claims benefit of U.S. Ser. No. 60/598,227,
filed Aug. 2, 2004.
TECHNICAL FIELD OF THE INVENTION
[0002] The invention relates to novel compounds which inhibit HIV
reverse transcriptase, a method for the treatment of HIV infection
using the compounds, and pharmaceutical compositions comprising the
compounds.
BACKGROUND OF THE INVENTION
[0003] The disease known as acquired immune deficiency syndrome
(AIDS) is caused by the human immunodeficiency virus (HIV),
particularly the strain known as HIV-1. In order for HIV to be
replicated by a host cell, the information of the viral genome must
be integrated into the host cell's DNA. However, HIV is a
retrovirus, meaning that its genetic information is in the form of
RNA. The HIV replication cycle therefore requires a step of
transcription of the viral genome (RNA) into DNA, which is the
reverse of the normal chain of events. An enzyme that has been
aptly dubbed reverse transcriptase (RT) accomplishes the
transcription of the viral RNA into DNA. The HIV virion includes
copies of RT along with the viral RNA.
[0004] Reverse transcriptase has three known enzymatic functions;
it acts as an RNA-dependent DNA polymerase, as a ribonuclease, and
as a DNA-dependent DNA polymerase. Acting as an RNA-dependent DNA
polymerase, RT transcribes a single-stranded DNA copy of the viral
RNA. Acting as a ribonuclease, RT destroys the original viral RNA,
and frees the DNA just produced from the original RNA. Finally,
acting as a DNA-dependent DNA polymerase, RT makes a second,
complementary DNA strand, using the first DNA strand as a template.
The two strands form double-stranded DNA, which is integrated into
the host cell's genome by another enzyme called integrase.
[0005] Compounds that inhibit the enzymatic functions of HIV-1
reverse transcriptase will inhibit replication of HIV-1 in infected
cells. Such compounds are useful in the prevention or treatment of
HIV-1 infection in human subjects, as demonstrated by known RT
inhibitors such as zidovudine, didanosine, zalcitabine, stavudine,
lamivudine, emtricitabine, abacavir, tenofovir, nevirapine,
delavirdine and efavirenz, the main reverse transcriptase
inhibitors thus far approved for use in the treatment of AIDS.
[0006] As with any antiviral therapy, use of RT inhibitors in the
treatment of AIDS eventually leads to a virus that is less
sensitive to the given drug. Resistance (reduced sensitivity) to
these drugs is the result of mutations that occur in the reverse
transcriptase segment of the pol gene. Several mutant strains of
HIV have been characterized, and resistance to known therapeutic
agents is believed to be due to mutations in the RT gene. One of
the more commonly observed mutants clinically for the
non-nucleoside reverse transcriptase inhibitors is the K103N
mutant, in which a lysine (K), at codon 103, has been mutated to a
asparagine (N) residue. Other mutants, which emerge with varying
frequency during treatment using known antivirals, include single
mutants Y181C, G190A, Y188C, and P236L, and double mutants
K103N/Y181C, K103N/P225H, K103N/V108I and K103N/L100I.
[0007] As antiviral use in therapy and prevention of HIV infection
continues, the emergence of new resistant strains is expected to
increase. There is therefore an ongoing need for new inhibitors of
RT, which have different patterns of effectiveness against the
various resistant mutants.
[0008] Antiviral entry inhibitors useful in treating HIV infection
have been described in WO 03/075907 (Tibotec) and non-nucleoside
inhibitors of HIV reverse transcriptase containing a benzophenone
moiety have been described in WO 01117982 (Glaxo) and WO 02/070470
(SmithKline Beecham). As well, non-nucleoside inhibitors of HIV
reverse transcriptase have been described in WO 2004/050643
(Boehringer Ingelheim).
[0009] The present invention provides novel compounds which show
potent-activity against wild type HIV reverse transcriptase as well
as against single mutant and double mutant strains.
SUMMARY OF THE INVENTION
[0010] The invention provides compounds of formula (I) which are
useful for treating HIV infection in a human infected by HIV. The
compounds are potent inhibitors of wild-type (WT) and double mutant
strains of HIV-1 RT, particularly the double mutant
K103N/Y181C.
[0011] In a first aspect the invention provides a compound,
represented by formula (I): ##STR2## wherein [0012] R.sup.1 and
R.sup.2 are each independently selected from H, halo, cyano,
(C.sub.1-4)alkyl, (C.sub.1-4)haloalkyl, (C.sub.2-4)alkenyl,
(C.sub.2-4)alkynyl and (C.sub.3-6)cycloalkyl; [0013] with the
proviso that when R.sup.1 is H, R.sup.2 cannot be H; [0014] R.sup.3
is halo; [0015] R.sup.4 is selected from (C.sub.1-4)alkyl, halo and
nitro; and [0016] R.sup.5 and R.sup.6 are each independently
selected from H, halo and (C.sub.1-4)alkyl; or a salt thereof.
[0017] According to a further aspect of the invention, there is
provided a pharmaceutical composition, comprising a compound of
formula (I) as defined hereinbefore and hereinafter, or a
pharmaceutically acceptable salt thereof, and optionally one or
more pharmaceutically acceptable carriers.
[0018] According to yet another aspect of the invention, there is
provided a pharmaceutical composition, comprising a compound of
formula (I) as defined hereinbefore and hereinafter, or a
pharmaceutically acceptable salt thereof, in combination with one
or more other antiretroviral drugs.
[0019] According to another aspect of the invention, there is
provided a pharmaceutical composition for the treatment of HIV
infection, comprising a compound of formula (I) as defined
hereinbefore and hereinafter, or a pharmaceutically acceptable salt
thereof, and optionally one or more pharmaceutically acceptable
carriers.
[0020] A further aspect of the invention provides a pharmaceutical
composition for the treatment of HIV infection, comprising a
compound of formula (I) as defined hereinbefore and hereinafter, or
a pharmaceutically acceptable salt thereof, and optionally one or
more pharmaceutically acceptable carriers, in combination with one
or more other antiretroviral drugs.
[0021] Another important aspect of the invention involves a method
of treating an HIV infection in a mammal by administering to the
mammal an anti-HIV effective amount of a compound of formula (I) as
defined hereinbefore and hereinafter, a pharmaceutically acceptable
salt thereof, or a composition as described above, alone or in
combination with at least one other antiretroviral agent,
administered together or separately.
[0022] Still another aspect of the invention provides the use of a
compound of formula (I) as defined hereinbefore and hereinafter, or
a pharmaceutically acceptable salt thereof, for the treatment of
HIV infection in a mammal.
[0023] According to another aspect of the invention, there is
provided a method of inhibiting HIV-1 replication by exposing the
virus to an inhibitory amount of a compound of formula (I) as
defined hereinbefore and hereinafter, or a pharmaceutically
acceptable salt thereof.
[0024] Yet another aspect of the invention provides the use of a
compound of formula (I) as defined hereinbefore and hereinafter, or
a pharmaceutically acceptable salt thereof, to inhibit
HIV-1-replication.
[0025] According to another aspect of the invention, there is
provided the use of a compound of formula (I) as defined
hereinbefore and hereinafter, or a pharmaceutically acceptable salt
thereof, for the manufacture of a medicament for the treatment of
an HIV infection.
[0026] According to yet another aspect of the invention, there is
provided the use of a compound of formula (I) as defined
hereinbefore and hereinafter, or a pharmaceutically acceptable salt
thereof, for the manufacture of a medicament for the treatment of
an HIV infection, in combination with one or more other
antiretroviral drugs.
[0027] Another aspect of the invention provides an article of
manufacture comprising a composition effective to treat an HIV
infection or to inhibit the reverse transcriptase of HIV; and
packaging material comprising a label which indicates that the
composition can be used to treat infection by the human
immunodeficiency virus; wherein the composition comprises a
compound of formula (I) as defined hereinbefore and hereinafter, or
a pharmaceutically acceptable salt thereof.
DETAILED DESCRIPTION OF THE INVENTION
Definitions
[0028] The following definitions apply unless otherwise noted:
[0029] As used herein, the term "(C.sub.1-n)alkyl" wherein n is an
integer, either alone or in combination with another radical, is
intended to mean acyclic straight or branched chain alkyl radicals
containing from one to n carbon atoms respectively. Examples of
such radicals include, but are not limited to, methyl (Me), ethyl
(Et), propyl (Pr), 1-methylethyl (iPr), butyl (Bu), 1-methylpropyl,
2-methylpropyl (iBu), and 1,1-dimethylethyl (tBu), wherein the
abbreviations commonly used herein are given in brackets.
[0030] As used herein, the terms "halo" or "halogen", used
interchangeably, mean a halo radical selected from bromo, chloro,
fluoro or iodo.
[0031] As used herein, the term "(C.sub.1-n)haloalkyl" wherein n is
an integer, means an alkyl radical containing one to n carbon atoms
wherein one or more hydrogen atoms are replaced by a halogen atom
(including but not limited to trifluoromethyl).
[0032] As used herein, the term "(C.sub.2-n)alkenyl" wherein n is
an integer, either alone or used with another radical, means an
unsaturated, acyclic, straight or branched chain radical containing
two to n carbon atoms, at least two of which are bonded to each
other by a double bond and includes, but is not limited to,
--CH.dbd.CH.sub.2, --CH.sub.2CH.dbd.CH.sub.2,
--CH.sub.2CH.dbd.CHCH.sub.3 and --CH(Me)CH.dbd.CH.sub.2. The cis
and trans isomers, and mixtures thereof, of the (C.sub.2-n)alkenyl
radical are encompassed by the term. A (C.sub.2-n)alkenyl radical
may be substituted on any of the carbon atoms thereof which would
otherwise bear a hydrogen atom.
[0033] The term "(C.sub.2-n)alkynyl", as used herein, wherein n is
an integer, either alone or in combination with another radical, is
intended to mean an unsaturated, acyclic straight or branched chain
radical containing two to n carbon atoms, at least two of which are
bonded to each other by a triple bond. Examples of such radicals
include, but are not limited to, ethynyl, 1-propynyl, 2-propynyl,
and 1-butynyl. A (C.sub.2-n)alkynyl radical may be substituted on
any of the carbon atoms thereof which would otherwise bear a
hydrogen atom.
[0034] The term "(C.sub.3-m)cycloalkyl" as used herein, wherein m
is an integer, either alone or in combination with another
substituent, means a cycloalkyl substituent containing from 3 to m
carbon atoms and includes, but is not limited to, cyclopropyl
(cPr), cyclobutyl (cBu), cyclopentyl, cyclohexyl and cycloheptyl,
wherein the abbreviations commonly used herein are given in
brackets.
[0035] As used herein, the term "inhibitor of HIV replication"
refers to an agent capable of reducing or eliminating the ability
of HIV-1 reverse transcriptase to replicate a DNA copy from an RNA
template.
[0036] As used herein, the term "single or double mutant strains"
means that either one or two amino acid residues that are present
in WT HIV-1 strain have been replaced by residues not found in the
WT strain. For example, for the single mutant Y181C, the tyrosine
at residue 181 of the wild type HIV reverse transcriptase enzyme
has been replaced by a cysteine residue. Similarly, for the double
mutant K103N/Y181C, an asparagine residue has replaced the lysine
at residue 103 of the wild type HIV reverse transcriptase enzyme
and a cysteine residue has replaced the tyrosine at residue
181.
[0037] The term "salt thereof" means any base addition salt of a
compound according to the invention; preferably a pharmaceutically
acceptable salt thereof.
[0038] As used herein, the term "pharmaceutically acceptable salt"
means a salt of a compound which is, within the scope of sound
medical judgment, suitable for use in contact with the tissues of
humans and lower animals without undue toxicity, irritation,
allergic response, and the like, commensurate with a reasonable
benefit/risk ratio, generally water or oil-soluble or dispersible,
and effective for their intended use. Where applicable and
compatible with the chemical properties of the compound of formula
(I), the term includes pharmaceutically-acceptable acid addition
salts and pharmaceutically-acceptable base addition salts. Lists of
suitable salts are found in, e.g., S. M. Birge et al., J. Pharm.
Sci., 1977, 66, pp. 1-19.
[0039] The term "pharmaceutically-acceptable base addition salt"
means those salts which retain the biological effectiveness and
properties of the free acids and which are not biologically or
otherwise undesirable, formed with inorganic bases such as ammonia
or hydroxide, carbonate, or bicarbonate of ammonium or a metal
cation such as sodium, potassium, lithium, calcium, magnesium,
iron, zinc, copper, manganese, aluminum, and the like. Particularly
preferred are the ammonium, potassium, sodium, calcium, and
magnesium salts. Salts derived from pharmaceutically-acceptable
organic nontoxic bases include salts of primary, secondary, and
tertiary amines, quaternary amine compounds, substituted amines
including naturally occurring substituted amines, cyclic amines and
basic ion-exchange resins, such as methylamine, dimethylamine,
trimethylamine, ethylamine, diethylamine, triethylamine,
isopropylamine, tripropylamine, tributylamine, ethanolamine,
diethanolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol,
dicyclohexylamine, lysine, arginine, histidine, caffeine,
hydrabamine, choline, betaine, ethylenediamine, glucosamine,
methylglucamine, theobromine, purines, piperazine, piperidine,
N-ethylpiperidine, tetramethylammonium compounds,
tetraethylammonium compounds, pyridine, N,N-dimethylaniline,
N-methylpiperidine, N-methylmorpholine, dicyclohexylamine,
dibenzylamine, N,N-dibenzylphenethylamine, 1-ephenamine,
N,N'-dibenzylethylenediamine, polyamine resins, and the like.
Particularly preferred organic nontoxic bases are isopropylamine,
diethylamine, ethanolamine, trimethylamine, dicyclohexylamine,
choline, and caffeine.
[0040] As used herein, the term "treatment" means the
administration of a compound or composition according to the
present invention to alleviate or eliminate symptoms of the HIV
disease and/or to reduce viral load in a patient. The term
"treatment" also encompasses the administration of a compound or
composition according to the present invention post-exposure of the
individual to the virus but before the appearance of symptoms of
the disease, and/or prior to the detection of the virus in the
blood, to prevent the appearance of symptoms of the disease and/or
to prevent the virus from reaching detectible levels in the blood,
and the administration of a compound or composition according to
the present invention to prevent perinatal transmission of HIV-1
from mother to baby, by administration to the mother before giving
birth and to the child within the first days of life
[0041] The following sign
is used in sub-formulas to indicate the bond which is connected to
the rest of the molecule as defined.
DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS
[0042] In the following preferred embodiments, groups and
substituents of the compounds of formula (I) according to this
invention are described in detail.
R.sup.1 and R.sup.2:
[0043] Preferably, R.sup.1 and R.sup.2 are each independently
selected from H, halo, cyano, (C.sub.1-4)alkyl,
(C.sub.1-4)haloalkyl, and (C.sub.3-6)cycloalkyl; [0044] with the
proviso that when R.sup.1 is H, R.sup.2 cannot be H.
[0045] More preferably, R.sup.1 and R2 are each independently
selected from H, fluoro, chloro, bromo, iodo, cyano, methyl,
trifluoromethyl and cyclopropyl; [0046] with the proviso that when
R.sup.1 is H, R.sup.2 cannot be H.
[0047] Most preferably, R.sup.1 and R.sup.2 are each independently
selected from H, fluoro, chloro, bromo, cyano, trifluoromethyl and
cyclopropyl; [0048] with the proviso that when R.sup.1 is H,
R.sup.2cannot be H.
[0049] Any and each individual definition of R.sup.1 and R.sup.2 as
set out herein may be combined with any and each individual
definition of R.sup.3, R.sup.4, R.sup.5 and R.sup.6as set out
herein.
R.sup.3:
[0050] Preferably, R.sup.3is chloro.
[0051] Any and each individual definition of R.sup.3 as set out
herein may be combined with any and each individual definition of
R.sup.1, R.sup.2, R.sup.4, R.sup.5 and R.sup.6 as set out
herein.
R.sup.4
[0052] Preferably, R.sup.4is selected from chloro, bromo, nitro and
methyl.
[0053] More preferably, R.sup.4is chloro, bromo or methyl.
[0054] Any and each individual definition of R.sup.4 as set out
herein may be combined with any and each individual definition of
R.sup.1, R.sup.2, R.sup.3, R.sup.5 and R.sup.6 as set out
herein.
R.sup.5:
[0055] Preferably, R.sup.5 is H, fluoro, chloro or methyl.
[0056] More preferably, R.sup.5 is H or fluoro.
[0057] Alternatively more preferably, R.sup.5 is fluoro, chloro or
methyl.
[0058] Any and each individual definition of R.sup.6 as set out
herein may be combined with any and each individual definition of
R.sup.1, R.sup.2, R.sup.3, R.sup.4 and R.sup.6 as set out
herein.
R.sup.6:
[0059] Preferably, R.sup.6 is H or fluoro.
[0060] More preferably, R.sup.6 is H.
[0061] Any and each individual definition of R.sup.6 as set out
herein may be combined with any and each individual definition of
R.sup.1, R.sup.2, R.sup.3, R.sup.4 and R.sup.5 as set out
herein.
[0062] In one embodiment is provided a compound of formula (I):
##STR3## wherein [0063] R.sup.1 and R.sup.2 are each independently
selected from H, halo, cyano, (C.sub.1-4)alkyl, (C.sub.2-4)alkenyl,
(C.sub.2-4)alkynyl and (C.sub.3-6)cycloalkyl; wherein said
(C.sub.1-4)alkyl is optionally substituted with one to three halo
substituents; [0064] with the proviso that when R.sup.1 is H,
R.sup.2 cannot be H;
[0065] R.sup.3 is halo;
[0066] R.sup.4 is selected from (C.sub.1-4)alkyl, halo and
nitro;
[0067] R.sup.5 is selected from H and halo; and
[0068] R.sup.6 is H;
[0069] or a pharmaceutically acceptable salt thereof.
[0070] A preferred embodiment of the invention provides a compound
of formula (I) wherein R.sup.1 and R.sup.2 are each independently
selected from H, halo, cyano, (C.sub.1-4)alkyl,
(C.sub.1-4)haloalkyl, and (C.sub.3-6)cycloalkyl; [0071] with the
proviso that when R.sup.1 is H, R.sup.2 cannot be H;
[0072] R.sup.3 is chloro;
[0073] R.sup.4 is chloro, bromo, nitro or methyl;
[0074] R.sup.5 is H, fluoro, chloro or methyl; and
[0075] R.sup.6 is H or fluoro.
[0076] A more preferred embodiment of the invention provides a
compound of formula (I) wherein
[0077] R.sup.1 and R.sup.2 are each independently selected from H,
fluoro, chloro, bromo, cyano, trifluoromethyl and cyclopropyl;
[0078] with the proviso that when R.sup.1 is H, R.sup.2 cannot be
H;
[0079] R.sup.3 is chloro;
[0080] R.sup.4 is chloro, bromo or methyl;
[0081] R.sup.5 is H, fluoro, chloro or methyl; and
[0082] R.sup.6is H or fluoro.
Specific Embodiments
[0083] Included within the scope of this invention is each single
compound of formula (I) as presented in Table 1.
[0084] The compounds of formula (I) are effective inhibitors of
wild type HIV as well as of the double mutant reverse transcriptase
enzyme K103NNY181C. The compounds of the invention may also inhibit
the single mutant enzymes V106A, Y188L, K103N, Y181C, P236L and
G190A (among others). The compounds may also inhibit other double
mutant enzymes including K103N/P225H, K103N/V108I and
K103N/L100I.
[0085] The compounds of formula (I) possess inhibitory activity
against HIV-1 replication. When administered in suitable dosage
forms, they are useful in the treatment of AIDS, ARC and related
disorders associated with HIV-1 infection. Another aspect of the
invention, therefore, is a method for treating HIV-1 infection
which comprises administering to a human being, infected by HIV-1,
a therapeutically effective amount of a compound of formula (I), as
described above. The compounds may also be used to prevent
perinatal transmission of HIV-1 from mother to baby, by
administration to the mother before giving birth and to the child
within the first days of life.
[0086] The compounds of formula (I) may be administered in single
or divided doses by the oral, parenteral or topical routes. A
suitable oral dosage for a compound of formula (I) would be in the
range of about 0.5 mg to about 3 g per day. A preferred oral dosage
for a compound of formula (I) would be in the range of about 100 mg
to about 800 mg per day for a patient weighing 70 kg. In parenteral
formulations, a suitable dosage unit may contain from about 0.1 to
about 250 mg of said compounds, preferably about 1 mg to about 200
mg, whereas for topical administration, formulations containing
about 0.01 to about 1% active ingredient are preferred. It should
be understood, however, that the dosage administration from patient
to patient would vary. The dosage for any particular patient will
depend upon the clinician's judgment, who will use as criteria for
fixing a proper dosage the size and condition of the patient as
well as the patient's response to the drug.
[0087] When the compounds of the present invention are to be
administered by the oral route, they may be administered as
medicaments in the form of pharmaceutical preparations that contain
them in association with a compatible pharmaceutical carrier
material. Such carrier material can be an inert organic or
inorganic carrier material suitable for oral administration.
Examples of such carrier materials include but are not limited to
water, gelatin, talc, starch, magnesium stearate, gum arabic,
vegetable oils, polyalkylene-glycols, petroleum jelly and the
like.
[0088] The compounds of formula (I) can be used in combination with
one or more other antiretroviral drug known to one skilled in the
art, as a combined preparation useful for simultaneous, separate or
sequential administration for treating HIV infection in an
individual. Examples of antiretroviral drugs, including approved
and investigational drugs, that may be used in combination therapy
with compounds of formula (I) include but are not limited to:
[0089] NRTIs (nucleoside or nucleotide reverse transcriptase
inhibitors; including but not limited to zidovudine, didanosine,
zalcitabine, stavudine, lamivudine, emtricitabine, abacavir, and
tenofovir); [0090] NNRTIs (non-nucleoside reverse transcriptase
inhibitors; including but not limited to nevirapine, delavirdine,
efavirenz, capravirine, etravirine, rilpivirine, GW695634 and
BILR.sup.355); [0091] protease inhibitors (including but not
limited to ritonavir, tipranavir, saquinavir, nelfinavir,
indinavir, amprenavir, fosamprenavir, atazanavir, lopinavir, VX-385
and TMC-114); [0092] entry inhibitors including but not limited to
CCR5 antagonists (including but not limited to maraviroc
(UK-427,857), SCH-417690, GW873140 and TAK-652), CXCR4 antagonists
(including but not limited to AMD-11070), fusion inhibitors
(including but not limited to enfuvirtide (T-20)) and others
(including but not limited to PRO-542 and BMS-488043); [0093]
integrase inhibitors (including but not limited to c-1605,
BMS-538158 and JTK-303); [0094] a TAT inhibitors; [0095] maturation
inhibitors (including but not limited to PA-457); [0096]
immunomodulating agents (including but not limited to levamisole);
and [0097] antifungal or antibacterial agents (including-but
not-limited-to fluconazole).
[0098] Moreover, a compound of formula (I) can be used with at
least one other compound of formula (I).
[0099] The pharmaceutical preparations can be prepared in a
conventional manner and finished dosage forms can be solid dosage
forms, including but not limited to tablets, dragees, capsules, and
the like, or liquid dosage forms, including but not limited to
solutions, suspensions, emulsions and the like. The pharmaceutical
preparations may be subjected to conventional pharmaceutical
operations such as sterilization. Further, the pharmaceutical
preparations may contain conventional adjuvants including but not
limited to preservatives, stabilizers, emulsifiers,
flavor-improvers, wetting agents, buffers, salts for varying the
osmotic pressure and the like. Solid carrier material which can be
used includes but is not limited to starch, lactose, mannitol,
methyl cellulose, microcrystalline cellulose, talc, silica, dibasic
calcium phosphate, and high molecular weight polymers (such as
polyethylene glycol).
[0100] For parenteral use, a compound of formula (I) can be
administered in an aqueous or non-aqueous solution, suspension or
emulsion in a pharmaceutically acceptable oil or a mixture of
liquids, which may contain bacteriostatic agents, antioxidants,
preservatives, buffers or other solutes to render the solution
isotonic with the blood, thickening agents, suspending agents or
other pharmaceutically acceptable additives. Additives of this type
include but are not limited to, tartrate, citrate and acetate
buffers, ethanol, propylene glycol, polyethylene glycol, complex
formers (including but not limited to EDTA), antioxidants
(including but not limited to sodium bisulfite, sodium
metabisulfite, and ascorbic acid), high molecular weight polymers
(including but not limited to liquid polyethylene oxides) for
viscosity regulation and polyethylene derivatives of sorbitol
anhydrides. Preservatives may also be added if necessary, including
but not limited to benzoic acid, methyl or propyl paraben,
benzalkonium chloride and other quaternary ammonium compounds.
[0101] The compounds of this invention may also be administered as
a solution for nasal application and may contain in addition to the
compounds of this invention suitable buffers, tonicity adjusters,
microbial preservatives, antioxidants and viscosity-increasing
agents in an aqueous vehicle. Examples of agents used to increase
viscosity include but are not limited to polyvinyl alcohol,
cellulose derivatives, polyvinylpyrrolidone, polysorbates or
glycerin. Microbial preservatives added include but are not limited
to benzalkonium chloride, thimerosal, chloro-butanol or phenylethyl
alcohol. Additionally, the compounds provided by the invention may
be administerable by suppository.
Methodology and Synthesis
[0102] In general, the compounds of formula (I) are prepared by
known methods from readily available starting materials, using
reaction conditions known to be suitable for the reactants. Scheme
1 illustrates the general methods used to prepare the compounds of
formula (I), wherein X is halo (e.g. Br or I), P is a protecting
group, and R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.5 and R.sup.6
are as defined herein. ##STR4##
[0103] Briefly, the aryllithium obtained by halogen-lithium
exchange from 1(ii) can be acylated with amide 1(i) to give
protected benzophenone 1(iii). Other organometallic derivatives can
also be used to acylate amide 1(i). Alternatively, protected
benzophenone 1(iii) can also be obtained from the aryllithium
derived from 1(iv) upon acylation with amide 1(v). R.sup.1 and
R.sup.2 substituents (e.g. Br) of intermediate 1(iii) can also be
converted to other R.sup.1 and R.sup.2 substituents (e.g. CN, cPr)
using methods known to one skilled in the art. Protected
benzophenone 1(iii) is deprotected to give the hydroxybenzophenone
1(vi) by methods known to those skilled in the art. For example,
the methyl ether of benzophenone 1(iii) (P.dbd.CH.sub.3) may be
demethylated conveniently by treatment with BBr.sub.3. The
hydroxybenzophenone 1(vi) can be O-alkylated with an
.alpha.-haloacetic acid ester in the presence of base to yield the
corresponding ether 1(vii) (P.dbd.--CH.sub.3, --CH.sub.2CH.sub.3 or
--C(CH.sub.3).sub.3, for example). Cleavage of the ester protecting
group using well-known conditions, such as aqueous base (in the
case where P.dbd.--CH.sub.3, or --CH.sub.2CH.sub.3) or
trifluoroacetic acid (in the case where P.dbd.--C(CH.sub.3).sub.3)
gives acid 1(viii). Alternatively, hydroxybenzophenone 1(vi) may be
transformed directly into acid 1(viii) by alkylation with an
a-haloacetic acid. This approach for the preparation of
benzophenone 1(viii) has been described by J. H. Chan et al. (J.
Med. Chem. 2004, 47, 1175-1182). Finally, the coupling of acid
1(viii) with aminobenzoic acid derivative 1(ix) using appropriate
activation of the acid (for example, transformation to the
corresponding intermediate acyl chloride) can provide compounds of
formula (I).
[0104] Scheme 2 illustrates a general method used to prepare
non-commercially available 4-aminobenzoic acids 1(ix), wherein P is
a protecting group, R.sup.4 is methyl or chloro and R.sup.5 and
R.sup.6 are as defined herein. ##STR5##
[0105] Briefly, aniline 2(i) can be transformed to iodo derivative
2(ii). Protection of the aniline by methods known to those of skill
in the art (such as, for example, transformation to the
corresponding acetamide wherein P.dbd.--C(.dbd.O)CH.sub.3) gives
2(iii). The aryllithium or arylmagnesium halide obtained by
iodine-metal exchange from 2(iii) can be transformed to 2(iv) by
trapping with carbon dioxide. Alternative methods to transform
2(iii) to acid 2(iv) are known to the skilled in the art. Finally,
removal of the protective group by well known methods such as
treatment with aqueous base gives 4-aminobenzoic acid 1(ix).
[0106] Scheme 3 illustrates an alternative method used to prepare
non-commercially available 4-aminobenzoic acids 1(ix), wherein P is
a protecting group such as --CH.sub.3 or --CH.sub.2CH.sub.3,
R.sup.4 is chloro or bromo, R.sup.5 is as defined herein and
R.sup.6 is H. ##STR6##
[0107] Briefly, aniline 3(i) can be transformed to 3(ii)
(R.sup.4.dbd.Cl or Br) by chlorination or bromination. Removal of
the protecting group by well known methods such as treatment with
aqueous base gives 4-aminobenzoic acid 1(ix).
EXAMPLES
[0108] The present invention is illustrated in further detail by
the following non-limiting examples. All reactions were performed
in a nitrogen or argon atmosphere unless otherwise stated. Room
temperature is from about 18.degree. C. to about 22.degree. C.
(degrees Celsius). Solution percentages or ratios express a volume
to volume relationship, unless stated otherwise. Purification by
reverse phase HPLC (RP-HPLC) was performed using a gradient of
MeCN/H.sub.2O containing TFA (0.06%) (CombiPrep ODS-AQ 50.times.20
mm, 5 .mu., 120 A). Analytical HPLC was carried out under standard
conditions using a Combiscreen ODS-AQ C18 reverse phase column,
YMC, 50.times.4.6 mm i.d., 5 .mu.M, 120 .ANG. at 220 nM, elution
with a linear gradient as described in the following table (Solvent
A is 0.06% TFA in H.sub.2O; solvent B is 0.06% TFA in CH.sub.3CN):
TABLE-US-00001 Time (min) Flow (mL/min) Solvent A (%) Solvent B (%)
0 3.0 95 5 0.5 3.0 95 5 6.0 3.0 50 50 10.5 3.5 0 100
Abbreviations or Symbols used herein Include: [0109] Ac: acetyl;
[0110] Bu: butyl; [0111] tBu: 1,1-dimethylethyl (tert-butyl);
[0112] cPr: cyclopropyl; [0113] Chaps:
3-{(3-cholamidopropyl)dimethylammonio}-1-propanesulfonate; [0114]
dGTP: deoxyguanosine triphosphate; [0115] DMF:
N,N-dimethylformamide; [0116] DMSO: dimethylsulfoxide; [0117] DTT:
DL-dithiothreitol; [0118] EDTA: ethylenediaminetetraacetic acid;
[0119] Et: ethyl; [0120] Et.sub.3N: triethylamine; [0121]
Et.sub.2O: diethyl ether; [0122] EtOH: ethanol; [0123] EtOAc: ethyl
acetate; [0124] GSH: glutathione; [0125] HPLC: high performance
liquid chromatography; [0126] iPr: 1-methylethyl (isopropyl);
[0127] Me: methyl; [0128] MeOH: methanol; [0129] MeCN:
acetonitrile; [0130] n-BuLi: n-butyllithium; [0131] NMR: nuclear
magnetic resonance; [0132] Ph: phenyl; [0133] Pr: propyl; [0134]
RP-HPLC: reverse phase high performance liquid chromatography;
[0135] TFA: trifluoroacetic acid; [0136] THF: tetrahydrofuran;
[0137] TLC: thin layer chromatography; [0138] Tris:
tris(hydroxymethyl)aminomethane. Syntheses
[0139] The following examples illustrate methods for preparing
compounds of the invention.
Example 1
Benzophenone Intermediate 1.6
[0140] ##STR7##
[0141] The modified method of J. H. Chan et al. (J. Med. Chem.
2004, 47, 1175-1182) was followed.
a) Compound 1.2
[0142] Acyl chloride 1.1 (5.00 g, 21.0 mmol) was added dropwise to
an ice-cold solution of MeNH(OMe).HCl (2.80 g, 28.1 mmol) and
Et.sub.3N (9.00 mL, 63.9 mmol) in CHCl.sub.3 (50 mL). The reaction
mixture was stirred at 0C for 45 min and at room temperature for 3
h then was concentrated under reduced pressure. The residue was
partitioned between water and EtOAc. The organic layer was washed
with water and brine, dried (MgSO.sub.4), filtered and concentrated
under reduced pressure to give compound 1.2 (4.78 g, 91% yield) as
a brown oil.
b) Compound 1.4
[0143] A solution of 2.5 M n-BuLi in hexane (28.7 mL, 71.7 mmol)
was added dropwise to a solution of compound 1.3 (16.0 g, 71.7
mmol) in Et.sub.2O (300 mL) at -78.degree. C. The reaction mixture
was stirred at -78.degree. C. for 20 min, then a solution of
compound 1.2 (18.0 g, 71.7 mmol) in Et.sub.2O (10 mL) was added
dropwise. The reaction mixture was stirred at -78.degree. C. for 30
min then was allowed to warm to room temperature and stirred at
room temperature for 30 min. The dark mixture was poured into water
and the mixture was extracted with EtOAc (2.times.). The organic
layers were washed with water and brine, dried (MgSO.sub.4),
filtered and concentrated under reduced pressure. The residue was
triturated with hexane to give 1.4 (13.3 g, 56% yield) as a white
solid.
c) Compound 1.5
[0144] To a cold (-78.degree. C.) solution of compound 1.4 (5.95 g,
17.9 mmol) in CH.sub.2Cl.sub.2 (100 mL) was added a solution of 1.0
M BBr.sub.3 in CH.sub.2Cl.sub.2 (50.0 mL, 50.0 mmol). The reaction
mixture was stirred at -78.degree. C. for 1 h then was allowed to
warm to room temperature and stirred at this temperature for 4 h.
The mixture was poured into ice-water and extracted with
CH.sub.2Cl.sub.2 (3.times.). The combined organic layers were
washed with water and brine, dried (MgSO.sub.4), filtered, and
concentrated under reduced pressure to give compound 1.5 (5.78 g,
100% yield) as a brown solid.
d) Compound 1.6
[0145] To a solution of phenol 1.5 (5.75 g, 18.0 mmol) in acetone
(40 mL) were added K.sub.2CO3 (10.0 g, 72.3 mmol) and ethyl
bromoacetate (2.20 mL, 19.4 mmol), and the mixture was heated at
reflux for 1 h. Upon cooling the reaction mixture was concentrated,
diluted with EtOAc and the resulting solution was successively
washed with water and brine, dried (MgSO.sub.4), filtered and
concentrated under reduced pressure. The residue was dissolved in a
mixture of THF (60 mL), EtOH (20 mL) and water (20 mL) and
LiOH.H.sub.2O (1.30 g, 31.0 mmol) was added. The solution was
stirred at room temperature for 3 days then was slowly acidified
with aqueous 1 N HCl solution. The mixture was concentrated under
reduced pressure to a volume of 30 mL and was partitioned between
water and EtOAc. The aqueous layer was extracted with EtOAc. The
combined organic layers were washed with water and brine, dried
(MgSO.sub.4), filtered and concentrated under reduced pressure. The
residue was allowed to crystallize from a mixture of Et.sub.2O and
hexane to give compound 1.6 (5.02 g, 75% yield) as a beige
solid.
Example 2
Benzophenone Intermediate 2.7
[0146] ##STR8## a) Compound 2.2
[0147] To a solution of acid 2.1 (20.3 g, 109 mmol) in
CH.sub.2Cl.sub.2 (500 mL) were added (COCl).sub.2 (14.0 mL, 157
mmol) and DMF (0.2 mL). After 2 h the reaction mixture was
concentrated under reduced pressure. A solution of the resulting
acyl chloride (22.3 g, 109 mmol) in CH.sub.2Cl.sub.2 (80 mL) was
added dropwise to a solution of Et.sub.3N (45.0 mL, 323 mmol) and
MeNH(OMe).HCl (13.9 g, 142 mmol) in CH.sub.2Cl.sub.2( 300 mL). The
resulting solution was stirred for 2 h at room temperature. The
reaction mixture diluted with EtOAc was successively washed with
aqueous 1 N HCl solution, aqueous saturated NaHCO.sub.3 solution
and brine, dried (MgSO.sub.4), filtered and concentrated under
reduced pressure to give compound 2.2 (24.3 g, 97% yield) as a
white solid.
b) Compound 2.4
[0148] Using a method similar to the one described in Example 1,
Step b, but starting with compound 2.3 (4.40 g, 17.3 mmol) and
compound. 2.2 (3.98 g, 17.3 mmol), compound 2.4 (3.60 g, 60% yield)
was obtained as a yellow solid.
c) Compound 2.5
[0149] A mixture of compound 2.4 (8.63 g, 25.1 mmol) and CuCN (6.75
g, 75.4 mmol, dried at 100.degree. C. under reduced pressure for 18
h) in DMF (50 mL) was heated at 185.degree. C. for 3.5 h. 20 The
cooled reaction mixture was diluted with EtOAc and the resulting
solution was washed with concentrated NH.sub.4OH solution, water
and brine, dried (MgSO.sub.4), filtered and concentrated to a
volume of about 50 mL. Hexane (150 m L) was then added and the
resulting precipitate was recovered by filtration and dried to give
compound 2.5 (5.70 g, 78% yield) as a off-white solid.
d) Compound 2.6
[0150] A solution of 1.0 M BBr.sub.3 in CH.sub.2Cl.sub.2(50.0 mL,
50.0 mmol) was added over 15 min to a cold (-78.degree. C.)
solution of compound 2.5 (5.70 g, 19.7 mmol) in CH.sub.2Cl.sub.2
(120 mL). The reaction mixture was stirred at -78.degree. C. for 1
h then was allowed to warm to room temperature (30 min). The
mixture was poured into ice-water. The phases were separated. The
aqueous layer was extracted with CH.sub.2Cl.sub.2 (3.times.). The
combined organic layers were washed with water, dried (MgSO.sub.4),
filtered and concentrated under reduced pressure. The residue was
dissolved in EtOAc (30 mL) and hexane (100 mL) was added. The
resulting green solid obtained by filtration was washed with hexane
(10 mL), dried under reduced pressure to give compound 2.6 (4.72 g,
87% yield).
e) Compound 2.7
[0151] A solution of phenol 2.6 (4.72 g, 17.1 mmol).
K.sub.2CO.sub.3 (7.09 g, 51.4 mmol) and t-butyl bromoacetate (2.82
mL, 17.5 mmol) in acetone (75 mL) was heated at 50.degree. C. for
1.5 h. The cooled reaction mixture diluted with EtOAc was washed
with water (2.times.) and brine, dried (MgSO.sub.4), filtered and
concentrated under reduced pressure. A solution of the residue in
TFA (25 mL) and CH.sub.2Cl.sub.2 (50 mL) was stirred at room
temperature for 1 h. The reaction mixture was concentrated under
reduced pressure to a volume of 40 mL, hexane (100 mL) was added
and the resulting suspension was filtered. The solid was washed
with hexane and dried under reduced pressure to give compound 2.7
(5.14 g, 90% yield) as a white solid.
Example 3
Benzophenone Intermediate 3.2
[0152] ##STR9## a) Compound 3.1
[0153] A solution of 1.6 M n-BuLi in hexane (7.2 mL, 11.5 mmol) was
added over 45 min to a cold (-78.degree. C.) solution of
cyclopropylbromide (1.17 mL, 14.5 mmol) in THF (40 mL). After 1 h,
a solution of ZnBr.sub.2 (flame dried under high vacuum, 2.88 g,
12.8 mmol) in THF (10 mL) was added by cannula and the mixture was
allowed to warm to room temperature. After 1 h a solution of
compound 2.4 (from Example 2) (2.00 g, 5.82 mmol) in THF (30 mL)
and Pd(PPh.sub.3).sub.4 (672 mg, 0.58 mmol, under stream of
nitrogen) were added. The reaction mixture was then heated at
reflux for 16 h, cooled in an ice bath and quenched with aqueous
saturated NaHCO.sub.3 solution. The resulting mixture was extracted
with EtOAc several times and the combined organic layers were
successively washed with water and brine, dried (MgSO.sub.4),
filtered and concentrated under reduced pressure. The crude product
was purified by flash chromatography (hexane/EtOAc, 90/10) to give
compound 3.1 (1.25 g, 70% yield) as a pale yellow solid.
b) Compound 3.2
[0154] Using a method similar to the one described in Example 1,
Steps c and d, but starting with compound 3.1 (1.20 g, 3.94 mmol),
acid 3.2 (1.30 g, 95% yield) was obtained as a white solid.
Example 4
Benzophenone Intermediate 4.1
[0155] ##STR10## a) Compound 4.1
[0156] Using a method similar to the one described in Example 1,
Steps c and d, but starting with compound 2.4 (2.00 g, 5.82 mmol),
compound 4.1 (1.16 g, 51% yield) was obtained as a beige solid.
[0157] Using the methods of Example 1 or Example 2, but starting
with commercially available appropriately substituted benzoic acid
or benzoyl chloride and bromobenzene intermediates, other
benzophenone intermediates used in the preparation of compounds of
formula (I) were prepared.
Example 5
Aniline Intermediate 5.5
[0158] ##STR11## a) Compound 5.2
[0159] To a solution of aniline 5.1 (5.20 g, 41.6 mmol) in acetic
acid (30 mL) was added Kl (8.46 g, 51 mmol), NaBO.sub.3.4H.sub.2O
(7.28 g, 47 mmol) and (NH.sub.4).sub.2MoO.sub.4 (7.9 g, 40 mmol).
After 30 min the reaction was poured into a mixture of aqueous
saturated NaHCO.sub.3 solution (50 mL) and aqueous 10%
Na.sub.2S.sub.2O.sub.3 solution (10 mL). The aqueous layer was
extracted with Et.sub.2O and the combined organic phases were
washed with brine, dried (MgSO.sub.4) filtered and concentrated
under reduced pressure to give compound 5.2 (9.96 g, 95% yield) as
a beige solid.
b) Compound 5.3
[0160] To a solution of aniline 5.2 (2.20 g, 8.76 mmol) in THF (25
mL) were added Et.sub.3N (1.40 mL, 10 mmol) and acetyl chloride
(0.64 mL, 9.0 mmol). After 3 h the reaction mixture was diluted
with Et.sub.2O and the resulting solution washed with aqueous 1 N
HCl solution, water and brine, dried (MgSO.sub.4) filtered and
concentrated under reduced pressure to give compound 5.3 (2.40 g,
93% yield) as a beige solid.
c) Compound 5.4
[0161] To a solution of 5.3 (0.30 g, 1.02 mmol) in THF (10 mL) was
added NaH (44.0 mg, 1.10 mmol). After 30 min the reaction was
cooled to -30.degree. C. and 2.0 M i-PrMgCl solution in THF (0.60
mL, 1.2 mmol) was added. The reaction mixture was allowed to warm
to room temperature and CO.sub.2(g) was bubbled through the
solution for 15 min. Upon quenching with aqueous 1 N HCl solution
(5 mL), the reaction mixture was extracted with EtOAc. The combined
organic phases were washed with brine, dried (MgSO.sub.4), filtered
and concentrated under reduced pressure. The resulting solid was
triturated with Et.sub.2O to give compound 5.4 (60 mg, 28% yield)
as a beige solid.
d) Compound 5.5
[0162] A solution of compound 5.4 (40 mg, 0.19 mmol) in aqueous 5 N
NaOH solution (3 mL) was heated at 80.degree. C. for 3 h. Upon
cooling the reaction mixture was acidified with aqueous 12 N HCl
solution and extracted with EtOAc. The combined organic phase were
washed with brine, dried (MgSO.sub.4), filtered and concentrated
under reduced pressure to give compound 5.5 (25 mg, 78% yield) as a
white solid.
Example 6
Aniline Intermediate 6.4
[0163] ##STR12## a) Compound 6.2
[0164] Following a procedure similar to the one described in
Example 5 steps a and b but starting with compound 6.1 (1.98 g,
14.0 mmol), compound 6.2 (2.70 g, 62% yield) was obtained as a grey
solid.
b) Compound 6.3
[0165] Following a procedure similar to the one described in
Example 5 step c but starting with compound 6.2 (1.00 g, 3.23
mmol), compound 6.3 (300 mg, 41% yield) was obtained as a white
solid.
c) Compound 6.4
[0166] Following a procedure similar to the one described in
Example 5 step d but starting with compound 6.3 (200 mg, 0.88
mmol), compound 6.4 (150 mg, 92% yield) was obtained as a white
solid.
Example 7
Aniline Intermediate 7.4
[0167] ##STR13## a) Compound 7.2
[0168] Following a procedure similar to the one described in
Example 5 steps a and b but starting with compound 7.1 (2.27 g,
14.0 mmol), compound 7.2 (2.80 g, 61% yield) was obtained as a
light pink solid.
b) Compound 7.3
[0169] Following a procedure similar to the one described in
Example 5 step c but starting with compound 7.2 (1.00 9, 3.03
mmol), compound 7.3 (420 mg, 56% yield) was obtained as a white
solid.
c) Compound 7.4
[0170] Following a procedure similar to the one described in
Example 5 step d but starting with compound 7.3 (250 mg, 1.01
mmol), compound 7.4 (180 mg, 87% yield) was obtained as a light
pink solid.
Example 8
Aniline Intermediate 8.3
[0171] ##STR14## a) Compound 8.2
[0172] To a solution of aniline 8.1 (1.00 g, 6.45 mmol) in MeCN (10
mL) was added N-chlorosuccinimide (860 mg, 6.45 mmol). The reaction
mixture was heated at 60.degree. C. for 3 h. After cooling to room
temperature the reaction mixture was concentrated under reduced
pressure. The residue was dissolved in CH.sub.2Cl.sub.2, treated
with an ethereal solution of diazomethane (.about.0.6 M, 20 mL) and
concentrated under reduced pressure. The residue was purified by
flash chromatography (hexane/EtOAc, 90/10 to 60/40) to give
compound 8.2 (280 mg, 21% yield) as a white solid.
b) Compound 8.3
[0173] To a solution of aniline 8.2 (29.5 mg, 145 .mu.mol) in THF
(5 mL) and MeOH (4 mL) was added an aqueous 1 N NaOH solution (4
mL). After 16 h aqueous 1 N HCl (25 mL) was added and the mixture
was extracted several times with EtOAc. The combined organic layers
were successively washed with water and brine, dried (MgSO.sub.4),
filtered and concentrated under reduced pressure to give compound
8.3 (25.4 mg, 92% yield) as a beige solid.
Example 9
Entry 1003
[0174] ##STR15## a) Compound 1003
[0175] To a solution of acid 2.7 (from Example 2) (39 mg, 0.12
mmol) in CH.sub.2Cl.sub.2 (2 mL) were added oxalyl chloride (11
.mu.L, 0.13 mmol) and DMF (1 drop). After 2 h the reaction was
concentrated under reduced pressure to give the corresponding acyl
chloride. Pyridine (12 .mu.L, 0.15 mmol) and compound 5.5 (from
Example 5) (20 mg, 0.12 mmol) were added to a solution of the crude
acyl chloride in THF (2 mL) at room temperature and the resulting
solution was stirred for 2 h. The reaction mixture was concentrated
under reduced pressure and the crude acid was purified by RP-HPLC.
The pure fractions were combined and concentrated to give compound
1003 (10 mg, 17% yield) as a white solid.
[0176] .sup.1H-NMR (DMSO-d.sub.6): .delta. 13.06 (broad s, 1H),
9.57 (s, 1H), 8.16 (m, 1H), 8.05 (s, 1H), 7.94 (m, 1H), 7.69 (dd,
J=9.0, 2.7 Hz, 1H), 7.66 (m, 1H), 7.56 (d, J =2.7 Hz, 1H), 7.45 (m,
1H), 7.24 (d, J=9.0 Hz, 1H), 4.82 (s, 2H), 2.04 (s, 3H).
Example 10
Entry 1001
[0177] ##STR16## a) Compound 10.1
[0178] To a solution of acid 1.6 (from Example 1) (150 mg, 0.398
mmol) in CH.sub.2Cl.sub.2 (10 mL) at room temperature were added
oxalyl chloride (104 .mu.L, 1.19 mmol) and DMF (1 drop). The
reaction mixture was stirred for 30 min then was concentrated under
reduced pressure to give acyl chloride 4.1 (157 mg, 100%
yield).
b) Compound 1001
[0179] Commercially available aniline 10.2 (120 mg, 0.796 mmol) and
pyridine (161 .mu.L, 1.99 mmol) were successively added to a
solution of crude acyl chloride 10.1 (157 mg, 0.398 mmol) in THF (1
0 mL) at room temperature. The reaction mixture was stirred at room
temperature for 1 h then was diluted with EtOAc. The resulting
solution was successively washed with aqueous 0.5 N HCl solution
(2.times.), water, aqueous saturated NaHCO.sub.3 solution and
brine, dried (MgSO.sub.4), filtered and concentrated under reduced
pressure. Half the crude residue was purified by RP-HPLC. The pure
fractions were combined and concentrated to give compound 1001 (44
mg, 43% yield) as a white solid. .sup.1H-NMR (DMSO-d.sub.6):
.delta. 12.81 (broad s, 1H), 9.35 (s, 1H), 8.01 (d, J=8.4 Hz, 1H),
7.89 (s, 1H), 7.88 (d, J=7.9 Hz, 1H), 7.77 (s, 1H), 7.72 (dd,
J=8.4, 1.4 Hz, 1H), 7.67 (dd, J=8.8, 2.5 Hz, 1H), 7.61 (d, J=8.4
Hz, 1H), 7.55 (d, J=2.8 Hz, 1H), 7.23 (d, J=9.0 Hz, 1H), 4.81 (s,
2H), 2.15 (s, 3H).
Example 11
Entry 1002
[0180] ##STR17## a) Compound 1002
[0181] Following a procedure similar to the one described in
Example 10, steps-a and b but starting with acid 1.6 (from Example
1) (71.0 mg, 188 .mu.mol) and aniline 5.5 (from Example 5) (32.5
mg, 188 .mu.mol), compound 1002 (49.0 mg, 49% yield) was obtained
as a white solid. .sup.1H-NMR (DMSO-d.sub.6): .delta. 13.05 (s,
1H), 9.54 (s, 1H), 8.01 (d, J=8.4 Hz, 1H), 7.87 (m, 2H), 7.65 (m,
2H), 7.55 (d, J=2.8 Hz, 1 H), 7.40 (d, J=9 Hz, 1H), 7.23 (d, J=9
Hz, 1H), 4.81 (s, 2H), 2.01 (s, 3H).
Example 12
Entry 1004
[0182] ##STR18## a) Compound 1004
[0183] Following a procedure similar to the one described in
Example 5, step e but starting with compound 3.2 (from Example 3)
(60.0 mg, 172 .mu.mol) and compound 10.2 (from Example 10) (26.5
mg, 346 .mu.mol), compound 1004 (13.3 mg, 16% yield) was obtained
as a white solid. .sup.1H-NMR (DMSO-d.sub.6): .delta. 12.19 (s,
1H), 9.19 (s, 1H), 7.77 (s, 1H), 7.74-7.61 (m, 3H), 7.45 (d, J=2.5
Hz, 1H), 7.38 (s, 1H), 7.27 (d, J=9.2 Hz, 1H), 7.22 (d, J=9.0 Hz,
1H), 7.15 (d, J=10 Hz, 1H), 4.81 (s, 2H), 2.15 (s, 3H), 2.04-1.99
(m, 1H), 0.98-0.93 (m, 2H), 0.71-0.67 (m, 2H).
Example 13
Reverse Transcriptase (RT) Assays
Enzymatic Assay (IC.sub.50)
[0184] The enzymatic assay employed is described as follows: The
reverse transcriptase (RT) enzyme assay has been adapted to a
96-well microtiter plate format and uses PicoGreen.TM. as a
fluorescent intercalator. More explicitly, the HIV-1 RT enzyme was
thawed and appropriately diluted into Tris/HCl 50 mM pH 7.8
containing NaCl 60 mM, MgCl.sub.2.6H.sub.2O 2 mM, DTT 6 mM, GSH 2
mM and 0.02% w/v Chaps to give .apprxeq.10 nM enzyme. To 10 .mu.L
of this enzyme solution was added 10 .mu.L of inhibitor solution
(40 .mu.M to 2.032 nM inhibitor in the same assay buffer as above
containing 4% v/v DMSO). The plate was pre-incubated for 15 minutes
at room temperature before proceeding to the next step. In this
pre-incubation step, the highest and lowest inhibitor
concentrations were 20 .mu.M and 1.016 nM respectively and the
concentration of DMSO was 2% v/v. Then the enzymatic reaction was
initiated by addition of 20 .mu.L of substrate solution. The final
reaction mixture contained Tris/HCl 50 mM pH 7.8, NaCl 60 mM,
MgCl.sub.2.6H.sub.2O 2 mM, DTT 6 mM, GSH 2 mM, CHAPS 0.02% w/v,
DMSO 1% v/v, poly rC 45 nM, dG.sub.15 4.5 nM, dGTP 3.6 .mu.M, and
.apprxeq.2.5 nM enzyme. In this incubation step, the highest and
lowest inhibitor concentrations were 10 .mu.M and 0.508 nM
respectively. After addition of the substrate cocktail, the plate
was covered with a plastic seal and incubated for 50 minutes at
37.degree. C. in a dry incubator. The reaction was then quenched by
addition of 5 .mu.L of EDTA 0.5 M. The plate was shaken for 30
seconds at medium speed and incubated for 5 minutes at room
temperature. Then 160 .mu.L of PicoGreen.TM. 1:400 dilution from
commercial stock (diluted in Tris 20mM pH 7.5 with EDTA 1 mM) was
added and the plate was shaken for 30 seconds and incubated for 10
minutes at room temperature. The plate was then analyzed using a
POLARstar Galaxy fluorometer (BMG Labtechnologies) with
.lamda..sub.ex and .lamda..sub.em of 485 nm and 520 nm
respectively. Each well was read for 1.25 second. Each row
contained at its extremities a blank and a control well.
P24 Cellular Assay (EC.sub.50)
[0185] The p24 assay is as described in WO 01/96338, pages 59-60,
herein incorporated by reference.
C8166 HIV-1 Luciferase Assay (EC.sub.50)
[0186] The luciferase assay is as described in WO 2004/050643,
pages 73-75, herein incorporated by reference.
Table
[0187] Table 1 illustrates further compounds of the present
invention, which can be synthesized in analogy to the methods as
described hereinbefore, optionally modified by procedures known to
the one skilled in the art. All compounds shown in the table are
active in at least one of the assays described in Example 13;
showing IC.sub.50 and/or EC.sub.50 values of less than 1 .mu.M.
[0188] Retention times (t.sub.R) for each compound were measured
using the standard analytical HPLC conditions described in the
Examples. As is well known to one skilled in the art, retention
time values are sensitive to the specific measurement conditions.
Therefore, even if identical conditions of solvent, flow rate,
linear gradient, and the like are used, the retention time values
may vary when measured, for example, on different HPLC instruments.
Even when measured on the same instrument, the values may vary when
measured, for example, using different individual HPLC columns, or,
when measured on the same instrument and the same individual
column, the values may vary, for example, between individual
measurements taken on different occasions. TABLE-US-00002 TABLE 1
##STR19## t.sub.R MS Cpd R.sup.1 R.sup.2 R.sup.4 R.sup.5 R.sup.6
(min) (MH.sup.+) 1001 F CF.sub.3 Me H H 6.9 510.1 512.1 1002 F
CF.sub.3 Me F H 7.0 526.1 528.0 (M - H).sup.- 1003 F CN Me F H 6.3
485.2 487.2 1004 F ##STR20## Me H H 7.1 482.2 484.2 1005 F Cl Me H
H 7.0 474.1 476.1 478.1 (M - H).sup.- 1006 H CN Me H H 7.1 447.1
449.1 (M - H).sup.- 1007 F CN Me H H 6.3 465.1 467.1 (M - H).sup.-
1008 F CF.sub.3 Cl H H 7.2 528.1 530.0 532.0 1009 F CN Cl H H 6.4
485.1 487.1 489.0 (M - H).sup.- 1010 F CF.sub.3 Cl F H 7.2 546.1
548.1 550.0 (M - H).sup.- 1011 F ##STR21## Me F H 7.0 500.1 502.1
1012 F Br Me F H 6.9 536.0 538.0 540.0 (M - H).sup.- 1013 F Cl Me F
H 6.9 492.0 494.0 496.0 (M - H).sup.- 1014 F Br Me H H 6.8 521.9
523.9 1015 H Br Me H H 5.9 500.0 502.0 504.0 (M - H).sup.- 1016 H
Br Me F H 6.7 521.8 523.9 1017 H CN Me F H 5.9 467.0 469.0 1018 Cl
CN Me H H 6.4 481.1 483.1 484.0 (M - H).sup.- 1019 F ##STR22## Br H
H 7.2 544.0 546.0 548.0 (M - H).sup.- 1020 F CF.sub.3 Br H H 7.1
572.0 573.9 576.0 (M - H).sup.- 1021 Cl CN Me F H 6.4 523.1 525.0
(M + Na).sup.+ 1022 F CF.sub.3 Cl Cl H 7.1 561.7 563.7 565.7 567.7
(M - H).sup.- 1023 F CF.sub.3 Me Me H 6.8 521.8 523.8 (M - H).sup.-
1024 F CF.sub.3 Me Cl H 6.9 543.8 545.8 1025 F CF.sub.3 Me H F 8.3
528.1 530.1
* * * * *